Ath 11 is complex: Raet1e and Esr1 as candidate genes; their mechanisms of action

Ath 11比较复杂：Raet1e和Esr1作为候选基因；

• 批准号: 8433465
• 负责人: JAN Leslie BRESLOW
• 金额: $ 50.17万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-04-01 至 2015-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8433465
• 关键词: Affect; Aorta; Arterial Fatty Streak; Atherosclerosis; Biological Assay; Breeding; Candidate Disease Gene; Cause of Death; Cells; Cellular Stress; Chromosomes, Human, Pair 10; Complex; Coronary heart disease; Data; Developing Countries; Distal; ESR1 gene; Early treatment; Estrogen Receptor 1; Estrogens; FVB/N Mouse; Female; Gender; Gene Cluster; Gene Expression; Gene Expression Microarray Analysis; Gene Expression Profile; Gene Expression Profiling; Genes; Genetic; Grant; Heritability; Histocompatibility Antigens Class I; Human; Immune; Immune system; Inbred Strain; Individual; Knockout Mice; Knowledge; Laboratories; Lesion; Ligands; MHC Class I Genes; Maps; Medical; Messenger RNA; Molecular; Mouse Strains; Mus; Myocardial Infarction; Nuclear Hormone Receptors; Pathway interactions; Physiological; Play; Population; Predisposition; Process; Proteins; Quantitative Trait Loci; Societies; Surface; Testing; Therapeutic; Transgenes; Transgenic Mice; Transgenic Organisms; Variant; Work; atherogenesis; atheroprotective; base; cell injury; congenic; cytokine; cytotoxicity; design; disability; genome wide association study; interest; macrophage; monocyte; mouse model; protein function; public health relevance; receptor; trait; transcription factor; tumor

DESCRIPTION (provided by applicant): Atherosclerosis is the major cause of death and disability in Westernized societies and is rapidly becoming so in developing nations. Despite significant medical progress our knowledge is incomplete and we still need to discover new genes and pathways in atherogenesis. Mouse models of atherosclerosis provide a complementary approach to human studies for such discoveries, and in an intercross between C57BL/6J and FVB/N mice on the apoEKO background we identified an atherosclerosis susceptibility locus on proximal mouse chromosome 10 (Ath11). Our work is at the forefront of the application of Quantitative Trait Locus (QTL) analysis not just to atherosclerosis but to all complex traits. We showed this locus contains 2 regions, a distal 10b region of 5 genes with Raet1e as a strong candidate gene, and a proximal 10a region of 21 genes with Esr1 an appealing candidate. The overall objective of this grant request is to confirm Raet1e and Esr1 as the culprit genes and elucidate their mechanisms of action. Raet1e is an MHC class I-like molecule expressed on the surface of stressed cells. It is a ligand for NKG2D, a receptor on the surface of NK, NKT, and T-Reg cells and some macrophages. This interaction plays a key role in the elimination of tumors and infected or damaged cells by the innate immune system. Our studies suggest aortic over expression of Raet1e is atheroprotective. To confirm this, we have made BAC Raet1e transgenic mouse strains and by breeding the transgene onto the apoEKO background we hope to prove our hypothesis. We will then identify the sequence variations between C57 and FVB in the Raet1e gene in the region of interest and find out which sequence variation is responsible. To determine how Raet1e over expression is atheroprotective we will analyze aortic gene expression patterns and cytokine profiles of mice with and without the Raet1e transgene. Using apoEKO and apoEKO NKG2DKO mice we will resolve if the effect of over expression of Raet1e on atherosclerosis is dependent or independent of the NKG2D receptor. We will identify by FACS analysis of splenic and aortic cells the immune cells involved in this process and with functional studies, such as cytotoxicity and proliferation assays, determine how over expression of Raet1e alters the physiological state of these cells. Esr1 is the main nuclear hormone receptor for estrogens. We will confirm Esr1 as the 10a region gene by expression analysis in subcongenic strains as well as comparing C57 and FVB Esr1 gene sequence, aortic mRNA and protein levels, and ESR1 function to identify the molecular and mechanistic basis of Esr1 as an atherosclerosis modifier gene. If we do not find support for Esr1 as a candidate gene for the 10a region we will identify, among the other 20 genes in the 10a region, the responsible gene by characterizing new subcongenic strains that further narrow the 10a region, and evaluate the most promising candidate genes by gene sequencing, gene expression analysis, and functional studies.

描述（由申请人提供）：动脉粥样硬化是西方社会死亡和残疾的主要原因，并且在发展中国家迅速变得如此。尽管医学取得了重大进展，但我们的知识还不完整，我们仍然需要发现动脉粥样硬化形成中的新基因和途径。动脉粥样硬化小鼠模型为此类发现提供了人类研究的补充方法，并且在 apoEKO 背景下的 C57BL/6J 和 FVB/N 小鼠之间的杂交中，我们在近端小鼠 10 号染色体 (Ath11) 上发现了动脉粥样硬化易感基因座。我们的工作处于定量性状基因座 (QTL) 分析应用的前沿，不仅适用于动脉粥样硬化，而且适用于所有复杂性状。我们表明该基因座包含 2 个区域，远端 10b 区域有 5 个基因，Raet1e 作为强候选基因，近端 10a 区域有 21 个基因，Esr1 是一个有吸引力的候选基因。该拨款申请的总体目标是确认 Raet1e 和 Esr1 为罪魁祸首基因并阐明其作用机制。 Raet1e 是一种在应激细胞表面表达的 MHC I 类分子。它是 NKG2D 的配体，NKG2D 是 NK、NKT 和 T-Reg 细胞以及一些巨噬细胞表面的受体。这种相互作用在先天免疫系统消除肿瘤和感染或受损细胞方面发挥着关键作用。我们的研究表明 Raet1e 的主动脉过度表达具有动脉粥样硬化保护作用。为了证实这一点，我们制备了 BAC Raet1e 转基因小鼠品系，并通过将转基因培育到 apoEKO 背景上，我们希望证明我们的假设。然后，我们将识别感兴趣区域 Raet1e 基因中 C57 和 FVB 之间的序列变异，并找出是哪个序列变异造成的。为了确定 Raet1e 过度表达如何具有动脉粥样硬化保护作用，我们将分析有和没有 Raet1e 转基因的小鼠的主动脉基因表达模式和细胞因子谱。使用apoEKO和apoEKO NKG2DKO小鼠，我们将确定Raet1e过度表达对动脉粥样硬化的影响是否依赖于或独立于NKG2D受体。我们将通过对脾细胞和主动脉细胞进行 FACS 分析来鉴定参与该过程的免疫细胞，并通过细胞毒性和增殖测定等功能研究来确定 Raet1e 的过度表达如何改变这些细胞的生理状态。 Esr1 是雌激素的主要核激素受体。我们将通过亚同类菌株的表达分析以及比较C57和FVB Esr1基因序列、主动脉mRNA和蛋白水平以及ESR1功能来确认Esr1为10a区基因，以确定Esr1作为动脉粥样硬化调节基因的分子和机制基础。如果我们没有找到支持 Esr1 作为 10a 区域候选基因的支持，我们将通过表征进一步缩小 10a 区域的新亚同类菌株，在 10a 区域的其他 20 个基因中确定负责基因，并评估最有希望的基因通过基因测序、基因表达分析和功能研究来候选基因。